Skip to main content
. 2020 Oct 14;21(12):e50642. doi: 10.15252/embr.202050642

Figure EV1. Merlin ubiquitination associates with activation of the Hippo pathway.

Figure EV1

  1. Total lysates from indicated cell lines cultured in regular conditions at a steady state were subjected to Western blotting. The same lysates were also used in Fig 1A.
  2. FH‐912 cells treated with DMSO or thapsigargin and subjected to Western blotting. The ratio of mono‐ubiquitinated to native Merlin in each lane was quantified by ImageJ and is shown under the blot. Native Merlin (asterisk); ubiquitinated Merlin (arrow).
  3. Met5‐A cells stably transduced with indicated shRNAs against Merlin or a scrambled shRNA were detached (Sus., suspension) by trypsinization using the procedure described in Materials and Methods and reseeded (Attach, attached) for 4 h. Total lysates from these cells were subjected to Western blotting.
  4. Longer exposure of the blot in Fig 1H. Ubiquitinated Merlin (arrows and arrowheads).
  5. LN229 cells transduced with vector or EGFP‐RAC1(Q61L) were treated with DMSO or thapsigargin (TG) and subjected to immunofluorescent staining. Scale bar = 20 μm.
  6. The ratio of the relative amount of YAP/TAZ in the nucleus (N) compared to the cytoplasm (C) from panel (E) was quantified. Mean ± s.e.m, two‐way ANOVA. ****P < 0.0001. Each data point represents one cell. N Vector = 16 cells in each condition, N RAC1(Q61L) = 17 cells in each condition. All cells were from multiple random images collected from one experiment. Two independent experiments were performed and gave similar results.

Source data are available online for this figure.